Cephalosporin derivatives and processes for the preparation thereof

ABSTRACT

The invention provides novel cephalosporin derivatives of the formula (I) ##STR1## and salts thereof for use in pharmaceutical compositions. Also novel precursors for synthesis of the cephalosporins are disclosed.

This application is the national phase under 35 U.S.C. §371 of prior PCTInternational Application No. PCT/KR96/00255 which has an internationalfiling date of Dec. 27, 1996 which designated the United States ofAmerica, the entire contents of which are hereby incorporated byreference.

FIELD OF THE INVENTION

The present invention relates to novel cephalosporin derivatives of theformula(I) and pharmaceutically acceptable non-toxic salts thereof; andmore particularly, to processes for preparing these compounds and topharmaceutical compositions containing the same as active ingredients,which have broad antibacterial activities against both Gram-positive andGram-negative bacteria.

SUMMARY OF THE INVENTION

An object of the present invention is to provide the novel cephalosporinderivatives of the formula(I) and pharmaceutically acceptable non-toxicsalts thereof. ##STR2## wherein R_(1a) is a hydrogen atom or an aminoprotecting group; R_(1b) is a hydrogen atom, C₁₋₄ alkyl group, C₂₋₄alkenyl group, C₂₋₄ alkynyl group, fluoro-substituted C₁₋₃ alkyl groupor a substituted or unsubstituted carboxyalkyl group (preferably methylgroup, ethyl group, allyl group, propargyl group, fluoromethyl group,2-fluoroethyl group, --C(CH₃)₂ COOH group, --CH₂ COOH group);

Q is CH or N;

R₂ and R₃ may be the same or different and mean individually a groupselected from C₁₋₃ alkyl group, C₂₋₄ alkenyl group, C₂₋₄ alkynyl groupand hydroxyl-substituted C₁₋₃ alkyl group; and

R₄ is ##STR3## wherein R₅ is a hydrogen atom, C₁₋₃ alkyl group or ahydroxyl-substituted C₁₋₃ alkyl group;

R₆ is a hydrogen atom, C₁₋₃ alkyl group, C₂₋₄ alkenyl group or an aminogroup;

R₇ is a C₁₋₃ alkyl group or a hydroxyl-substituted C₁₋₃ alkyl group;

Y is O or S or N; and

Z is a hydrogen atom, C₁₋₃ alkyl group, chlorine or fluorine atom.

DETAILED DESCRIPTION OF THE INVENTION

The cephalosporin compounds of the formula(I) can be prepared by thedisplacement reaction of the compounds of the formula(III) with thecompounds of the formula(IV).

The compounds of the formula(III) can be prepared in accordance withknown methods(J. of. Antibiotics., 43, 5, 533, 1990. European Patent No.0333154). ##STR4## wherein R_(1a), R_(1b), R₂, R₃, R₄ and Q are the sameas defined above;

R₈ is a hydrogen atom or a carboxyl protecting group; and

L is a leaving group.

Also, the cephalosporin compounds of the formula(I) can be prepared bythe acylation reaction of the compounds of the formula(VIII) with theactivated form of the compounds of the formula(IX).

The compounds of the formula(IX) can be prepared in accordance withknown methods (J. of. Antibiotics., 36, 8, 1020, 1983. J. of.Antibiotics., 45, 5, 709, 1992). ##STR5## wherein R_(1a), R_(1b), R₂,R₃, R₄ R₈, Q and L are the same as defined above;

R₉ is ani acyl group, formyl group, salicylaldehyde group orbenzaldehyde group; and

X is a halogen atom or an acid residue.

Specific examples of the compounds of formula(I) provided by thisinvention are shown below:

7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

7β-[(Z)-2-(2-aminothiazol-4-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

7β-[(Z)-2-(2-aminothiazol-4-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

7β-[(Z)-2-(5-amnino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateor pharmaceutically acceptable non-toxic salts thereof.

The following preparations and examples are provided for the purpose ofillustrating certain aspects of the present invention only; and are notto be construed as limiting the scope of the present invention in anyway.

Preparation 1

Preparation of 5-dimethylaminomethyl-4-methyloxazole

a) Preparation of 4-methyl-5-oxazolecarboxylic acid ethyl ester

To ethyl-2-chloroacetoacetate(5.6 g) was added formamide(4.6 g). Thereaction mixture was stirred at 120° C. for 12 hours, cooled to 0° C.and aqueous potassium carbonate was added thereto. The mixture wasextracted with benzene, dried with anhydrous magnesium sulfate, andconcentrated in vacuo to obtain the target product(2.1 g).

NMR(CDCl₃) δ (ppm): 1.40(t,3H), 2.50(s,3H), 4.35(q,2H), 7.84(s,1H).

b) Preparation of 5-hydroxymethyl-4-methyloxazole

After 4-methyl-5-oxazolecarboxylic acid ethyl ester(3 g) was dissolvedin ethyl alcohol(50 ml), sodiumborohydride(3 g) was added thereto andrefluxed for 17 hours. The reaction mixture was concentrated underreduced pressure, water was added to the concentrated solution. Afterextraction with ethyl acetate dried with anhydrous magnesium sulfate,and then concentrated to obtain the target product(0.9 g).

NMR(CDCl₃) δ (ppm): 2.18(s,3H), 3.45(s,1H), 4.65(s,2H), 7.80(s,1H).

c) Preparation of 5-dimethylaminomethyl-4-methyloxazole

To a solution of 5-hydroxymethyl-4-methyloxazole(2 g) dissolved indichloromethane(20 ml) were added thionylchloride(3.9 ml) at 0° C. Afterstirring at room temperature for 4 hours, ice-water(15 ml) were addedthereto. The seperated organic layer was dehydrated and concentrated.The residue were added ethylalcohol(25 ml) dimethylamine hydrochloride(2.5 g) and potassium carbonate(6.3 g). After stirring at roomtemperature for 4 hours, the solid was filtered off. The filtrate wasconcentrated under reduced pressure, water and dichloromethane addedthereto. The separated organic layer was dehydrated, and concentrated toobtain the target product(0.9 g).

NMR(CDCl₃) δ (ppm): 2.12(s,3H), 2.24(s,6H), 3.42(s,2H), 7.78(s,1H).

Preparation 2

Preparation of 5-dimethylaminomethyl-4-methylthiazole

a) Preparation of 4-methyl-5-thiazolecarboaldehyde.

To a solution of pyridinium dicromate(164.19 g) in dichloromethane (600ml), water(20 ml) was added. A solution of 4-methyl-5-thiazoleethanol(10g) in dichloromethane(500 ml) was added dropwise to the mixture and theresulting mixture stirred at room temperature for 24 hours. The solidwas filtered off and washed with water. The separated organic layer wasdehydrated, and concentrated to obtain the target product(6 g).

NMR(CDCl₃) δ (ppm): 2.79(s,3H), 9.02(s,1H), 10.18(s,1H).

b) Preparation of 4-methyl-5-thiazolemethanol

To sodiumborohydride(2.32 g) was added tetrahydrofurane(35 ml)at 0° C. Asolution of 4-methyl-5-thiazolecarboaldehyde(6 g) in tetrahydrofurane(35 ml) was added dropwise to the mixture and the resulting mixturestirred at room temperature for 3 hours. To the reaction mixture wasadded ice-water(60 ml) and saturated brine(30 ml). The separated organiclayer was dehydrated, and concentrated to obtain the target product(5g).

NMR(CDCl₃) δ (ppm): 2.35(s,3H), 4.79(s,2H), 8.61(s,1H).

c) Preparation of 5-dimethylaminomethyl-4-methylthiazole

To a solution of 4-methyl-5-thiazolemethanol(1.5 g) in chloroform(50ml), thionylchloride (1.7 ml) was added, and the mixture was refluxedfor 2 hours. The reaction mixture was concentrated under reducedpressure. The residue were added ethylalcohol(25 ml), dimethylaminehydrochloride (1.33 g) and potassium carbonate(2.3 g). After refluxingfor 5 hours, the solid was filtered off. The filtrate was concentratedunder reduced pressure, water and chloroform added thereto. Theseparated organic layer was dehydrated, and concentrated to obtain thetarget product(1 g).

NMR(CDCl₃) δ (ppm): 2.24(s,6H), 2.41(s,3H), 3.63(s,2H), 8,62(s,1H).

Preparation 3

Preparation of 4-ethylmethylamino-2-butenylamide

a) Preparation of 4-bromo-2-butenenitrile

To a solution of allylcyanide(40 g) dissolved in tert-butanol(60 ml) andpetroleum ether(273 ml) was added bromine(30.6 ml) dissolved intert-butanol(60.8 ml) at 15° C., and then stirred at room temperaturefor 15˜30 minutes. To sodium ethoxide(222.6 ml, 21%) was added dropwiseto the mixture and the solid was filtered off. The filtrate wasdistilled under reduced pressure to obtain the target product(55 g) b.p:80˜85° C.(12 mm).

NMR(CDCl₃) δ (ppm): 4.10(s,2H), 5.54(m,1H), 6.81(m,1H).

b) Preparation of 4-bromo-2-butenylamide

To a solution of 4-bromo-2-butenenitrile(20 g) suspended in water (2.3ml) was added slowly sulfuric acid(7.8 ml) at 40° C., and then stirredat 80˜90° C. for an hour. The reaction mixture was cooled to 40° C., andthen added ice(40 g) and ammonia solution(19.8 ml). The resultingprecipitate was collected by filtration and recrystallized fromethylacetate to obtain the target product(7.7 g).

NMR(CDCl₃) δ (ppm): 4.15(d,2H), 5.8˜6.2(br m,1H, NH₂), 6.92(m,1H).

c) Preparation of 4-ethylmethylamino-2-butenylamide

To a solution of 4-bromo-2-butenylamide(5.4 g) in acetonitrile(55 ml),N-ethylmethylamine(5.7 ml) was added at 0° C. The reaction mixture wasstirred for an hour and the resulting precipitate was collected byfiltration to obtain the target product(3.1 g).

NMR(DMSO-d₆) δ (ppm): 1.01(t,2H), 2.13(s,3H), 2.35(q,2H), 3.04(d,2H),5.99(d,1H), 6.55(m,1H).

EXAMPLE 1 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

To a solution of p-methoxybenzyl7β-[(Z)-2-methoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]-3-[(Z)-3-chloro-1-propen-1-yl]3-cephem-4-carboxylate(1.5g) dissolved in acetone(30 ml), sodium iodide(0.82 g) was added theretounder ice-cooling. The resulting solution was stirred for 15 minutesunder ice-cooling and for additional 60 minutes at room temperature. Thesolvent was distilled off, and the residue was extracted with ethylacetate. The extract was washed with a 10%-sodium thiosulfate and with asaturated brine, followed by drying with addition of anhydrous magnesiumsulfate. The dried extract was concentrated under reduced pressure andthe resulting precipitates were collected by filtration to obtain 1.2 gof p-methoxybenzyl7β-[(Z)-2-methoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]-3-[(E)-3-iodo-1-propen-1-yl]3-cephem-4-carboxylate.The powder was dissolved in N,N-dimethylformamide(2 ml), and5-dimethylaminomethyl-4-methylthiazole(0.31 g) was added thereto at -10°C. The resulting solution was stirred for 4 hours at -10° C. 10%-sodiumthiosulfate(70 ml) was added to the reaction solution, and the resultingprecipitates were collected by filtration to obtain the yellowish brownpowder. The powder was stirred in 88% formic acid(1.8 ml) for an hour at40° C., and acetone(400 ml) was added thereto. The resultingprecipitates were collected by filtration and washed with acetone. Theprecipitates were chromatographed over silicagel [acetonitrile:distilledwater 4:1(v/v)], Sephadex LH-20(or Amberlite XAD-2), and thenlyophilized to obtain the target product (250 mg) as white solid.

mp: 181° C.(dec.); IR(KBr,cm⁻¹): 1762, 1663, 1608, 1530; NMR(DMSO-d₆) δ(ppm): 2.43(s,3H), 2.94(s,6H), 3.42(s,2H), 3.82(s,3H), 4.01(d,2H),4.62(s,2H), 5.04(d,1H), 5.58(d,1H), 5.62(br d,1H), 7.1˜7.4(d,1H. s,1H.s,1H), 9.62(d,1H)

EXAMPLE 2 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(5-methylimidazol-4-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

The same procedures as described in Example 1 were repeated using 0.17 gof 4-dimethylaminomethyl-5-methylimidazole in place of5-dimethylaminomethyl-4-methylthiazole to obtain the target product(100mg).

mp: 178° C.(dec.); IR(KBr,cm⁻¹): 1760, 1670, 1610, 1531; NMR(DMSO-d₆) δ(ppm): 2.25(s,3H), 2.98(s,6H), 3.38(s,2H), 3.80(s,3H), 4.11(d,2H),4.59(s,2H), 5.01(d,1H), 5.53(d,1H), 5.59(br d,1H), 7.2˜7.84(d,1H. s,1H.s,1H), 9.66(d,1H).

EXAMPLE 3 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(2-aminothiazol-4-yl)-methyldimethylammonio]-1-propen-1-yl]-3-cephem-4-carboxylate

The same procedures as described in Example 1 were repeated using 0.21 gof 4-dimethylaminomethyl-2-aminothiazole in place of5-dimethylaminomethyl-4-methylthiazole to obtain the target product(150mg).

mp: 192° C.(dec.); IR(KBr,cm⁻¹): 1760, 1672, 1600, 1530; NMR(DMSO-d₆) δ(ppm): 2.12(s,6H), 3.35(s,2H), 3.83(s,3H), 4.01(d,2H), 4.55(s,2H),5.02(d,1H), 5.58(d,1H), 5.62(br d,1H), 6.72(br,2H), 7.1˜7.5(d,1H. s,1H.s,1H), 9.60(d,1H).

EXAMPLE 4 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(thiopen-2-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

The same procedures as described in Example 1 were repeated using 0.18 gof 2-thiopenmethyldimethylamine in place of5-dimethylaminomethyl-4-methylthiazole to obtain the target product (170mg).

mp: 186° C.(dec.); IR(KBr,cm⁻¹): 1762, 1670, 1600, 1530; NMR(DMSO-d₆) δ(ppm): 2.98(s,6H), 3.32(s,2H), 3.88(s,3H), 4.01(d,2H), 4.49(s,2H),5.02(d,1H), 5.55(d,1H), 5.62(br d,1H), 6.70(br d,2H. 1H), 7.1˜7.7(d,1H.s,1H. s,1H), 9.60(d,1H).

EXAMPLE 5 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(4-methyloxazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

The same procedures as described in Example 1 were repeated using 0.31 gof 5-dimethylaminomethyl-4-methyloxazole in place of5-dimethylaminomethyl-4-methylthiazole to obtain the target product (300mg).

mp: 199° C.(dec.); IR(KBr,cm⁻¹): 1762, 1668, 1610, 1533; NMR(DMSO-d₆) δ(ppm): 2.25(s,2H), 2.95(s,6H), 3.42(s,2H), 3.84(s,1H), 4.01(d,2H),4.62(s,2H), 5.04(d,1H), 5.58(d,1H), 5.62(br d,1H), 7.1˜7.3(d,1H. s,1lH.s,1H), 9.58(d,1H).

EXAMPLE 6 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

The same procedures as described in Example 1 were repeated using 0.23 gof 4-ethylmethylamino-2-butenylamide in place of5-dimethylaminomethyl-4-methylthiazole to obtain the target product (190mg).

mp: 189° C.(dec.); IR(KBr,cm⁻¹): 1761, 1675, 1608, 1530; NMR(DMSO-d₆) δ(ppm): 1.29(t,3H), 2.98(s,3H), 3.82(s,3H), 3.85(q,2H), 3.90(d,2H),3.94(d,2H), 4.75(s,2H), 5.08(d,1H), 5.62(d,1H), 5.68(br d,1H),6.75(d,1H), 6.99-7.12(br d,1H), 7.21˜7.38(d,1H. s,1H), 9.59(d,1H).

EXAMPLE 7 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-dimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

The same procedures as described in Example 1 were repeated using 0.25 gof 4-dimethylamino-2-butenylamide in place of5-dimethylaminomethyl-4-methylthiazole to obtain the target product(200mg).

mp: 198° C.(dec.); IR(KBr,cm⁻¹): 1763, 1670, 1600, 1530; NMR(DMSO-d₆) δ(ppm): 3.11(s,6H), 3.88(s,3H), 3.92(d,2H), 3.95(d,2H), 4.07(s,2H),5.05(d,1H), 5.62(d,1H), 5.69(br d,1H), 6.73(d,1H), 7.01-7.14(br d,1H),7.18˜7.32(d,1H. s,1H), 9.57(d,1H).

EXAMPLE 8 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methylethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

The same procedures as described in Example 1 were repeated using 0.35 gof 5-ethylmethylaminomethyl-4-methylthiazole in place of5-dimethylaminomethyl-4-methylthiazole to obtain the target product (300mg).

mp: 194° C.(dec.); IR(KBr,cm⁻¹): 1762, 1662, 1610, 1530; NMR(DMSO-d6) δ(ppm): 1.33(t,3H), 2.44(s,3H), 2.92(s,3H), 3.43(s,2H), 3.85(q,2H),3.87(s,3H), 4.01(d,2H), 4.60(s,2H), 5.05(d,1H), 5.56(d,1H), 5.67(brd,1H), 7.18˜7.32(d,1H. s,1H. s,1H), 9.60(d,1H).

EXAMPLE 9 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oxyimino)acetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

p-methoxybenzyl7β-[(Z)-2-(2-carboxyprop-2-oxyimino)-2-(2-tritylaminothiazol-4-yl)acetamido]-3-[(Z)-3-chloro-1-propen-1-yl]3-cephem-4-carboxylate(1.5g), sodium iodide (0.75 g) and 4-ethylmethylamino-2-butenylamide (0.28g) were reacted in the same manner as described in Example 1 to obtainthe target product(210 mg).

mp: 204° C.(dec.); IR(KBr, cm-1): 1760, 1663, 1610, 1530. NMR(DMSO-d₆) δ(ppm): 1.29(t,1H), 1.49(s,6H), 2.98(s,3H), 3.85(q,2H), 3.90(d,2H),3.94(d,2H), 4.75(s,2H), 5.08(d,1H), 5.62(d,1H), 5.68(br d,1H),6.75(d,1H), 6.99˜7.12(br d,1H), 7.21˜7.38(d,1H. s,1H), 9.59(d,1H).

EXAMPLE 10 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oxyimino)acetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

p-methoxybenzyl7β-[(Z)-2-(2-carboxyprop-2-oxyimino)-2-(2-tritylaminothiazol-4-yl)acetamido]-3-[(Z)-3-chloro-1-propen-1-yl]3-cephem-4-carboxylate(1.5g), sodium iodide (0.75 g) and5-dimethylam-inomethyl-4-methylthiazole(0.31 g) were reacted in the samemanner as described in Example 1 to obtain the target product(180 mg).

mp: 198° C. (dec.); IR(KBR, cm-1): 1762, 1665, 1610, 1525. NMR(DMSO-d₆)δ (ppm): 1.50(s,6H), 2.43(s,3H), 2.91(s,6H), 3.41(s,2H), 4.03(d,2H),4.60(d,2H), 4.60(s,2H), 5.05(d,1H), 5.57(d,1H), 5.65(br d,1H),7.1˜7.4(d,1H. s,1H. s,1H), 9.62(d,1H).

EXAMPLE 11 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

p-methoxybenzyl7β[(Z)-2-fluoroethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]3-[(Z)-3-chloro-1-propen-1-yl]3-cephem-4-carboxylate(1.3 g), sodium iodide(0.68 g) and5-dimethylaminomethyl-4-methylthiazole(0.27 g) were reacted in the samemanner as described in Example 1 to obtain the target product (300 mg).

mp: 196° C.(dec.); IR(KBr,cm⁻¹): 1765, 1665, 1610, 1530; NMR(DMSO-d₆) δ(ppm): 2.41(s,3H), 2.95(s,6H), 3.42(s,2H), 4.04(d,2H), 4.25(t,2H),4.53(t,2H), 4.79(s,2H), 5.04(d,1H), 5.56(d,1H), 5.61(br d,1H),7.1˜7.4(d,1H. s,1H. s,1H), 9.65(d,1H).

EXAMPLE 12 Synthesis of7β-[(Z)-2-(2-aminothiazol-4-yl)-2-fluoroethoxyiminoacetamdo]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

p-methoxybenzyl7β-[(Z)-2-fluoroethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]-3-[(Z)-3-chloro-1-propen-1-yl]3-cephem-4-carboxylate(1.3 g), sodium iodide(0.68 g) and 4-ethylmethylamino-2-butenylamnide(0.25 g) were reacted in the same manner as described in Example 1 toobtain the target product(230 mg).

mp: 191 ° C.(dec.); IR(KBr,cm⁻¹): 1760, 1672, 1600, 1530; NMR(DMSO-d₆) δ(ppm): 1.27(t,1H), 2.99(s,3H), 3.85(q,2H), 3.90(d,2H), 3.94(d,2H),4.27(t,2H), 4.52(t,2H), 4.75(s,2H), 5.09(d,1H), 5.60(d,1H), 5.69(brd,1H), 6.74(d,1H), 7.19˜7.35(d,1H. s,1H), 9.61(d,1H).

EXAMPLE 13 Synthesis of7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

To a solution of p-methoxybenzyl7β[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[(Z)-3-chloro-1-propen-1-yl]3-cephem-4-carboxylate(1.5g) dissolved in acetone(20 ml), sodium iodide (0.82 g) was added theretounder ice-cooling. The resulting solution was stirred for 15 minutesunder ice-cooling and for additional an hour at room temperature. Thesolvent was distilled off, and the residue was extracted with ethylacetate. The extract was washed with a 10%-sodium thiosulfate and with asaturated brine, followed by drying with addition of anhydrous magnesiumsulfate. The dried extract was concentrated under reduced pressure andthe resulting precipitates were collected by filtration to obtain 1.1 gof p-methoxybenzyl7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[(E)-3-iodo-1-propen-1-yl]3-cephem-4-carboxylate.The powder was dissolved in N,N-dimethylformamide(2.5 ml), and4-ethylmethylamino-2-butenylamide(0.2 g) was added thereto at -10° C.The resulting solution was stirred for 4 hours at -10° C. 10%-sodiumthiosulfate(70 ml) was added to the reaction solution, and the resultingprecipitates were collected by filtration to obtain the yellowish brownpowder. The powder was stirred in 88%-formic acid(1.3 ml) for 3 hours atroom temperature, and acetone(400 ml) was added thereto. The resultingprecipitates were collected by filtration and washed with acetone. Theprecipitates were chromatographed over silicagel[acetonitrile:distilledwater 4:1 (v/v)], Sephadex LH-20(or Amberlite XAD-2), and thenlyophilized to obtain the target product (180 mg) as white solid.

mp: 202° C.(dec.); IR(KBr,cm⁻¹): 1765, 1670, 1600, 1520; NMR(DMSO-d₆) δ(ppm): 1.29(t,3H), 2.99(s,3H), 3.83(s,3H), 3.84(q,2H), 3.91(d,2H),3.94(d,2H), 4.73(s,2H), 5.07(d,1H), 5.66(d,1H), 5.69(br d,1H),6.59(dd,1H), 6.99˜7.15(br d,1H), 7.28(dd, 1H), 8.63(d,1H), 9.59(d,1H).

EXAMPLE 14 Synthesis of7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(4-methylthiazol-5-yl)-methyldimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

p-methoxybenzyl7β[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-iodo-1-propen-1-yl]3-cephem-4-carboxylate(1.1g) and 5-dimethylaminomethyl-4-methylthiazol (0.36 g) were reacted inthe same manner as discribed in Example 13 to obtain the targetproduct(330 mg).

mp: 188° C.(dec.); IR(KBr,cm⁻¹): 1763, 1670, 1595, 1522; NMR(DMSO-d₆) δ(ppm): 2.40(s,3H), 2.93(s,6H), 3.42(s,2H), 4.04(d,2H), 4.28(t,2H),4.55(t,2H), 4.78(s,2H), 5.03(d,1H), 5.57(d,1H), 5.61(br d,1H),7.1˜7.32(d,1H. s,1H), 9.63(d,1H).

EXAMPLE 15 Synthesis of7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

p-methoxybenzyl7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-iodo-1-propen-1-yl]3-cephem-4-carboxylate(1g) and 4-ethylmethylamino-2-butenylamide(0.3 g) were reacted in the samemanner as described in Example 13 to obtain the target product(290 mg).

mp: 196° C.(dec.); IR(KBr,cm⁻¹): 1762, 1680, 1600, 1520; NMR(DMSO-d₆) δ(ppm): 1.27(s,1H), 2.98(s,3H), 3.83(q,2H), 3.91(d,2H), 3.94(d,2H),4.27(t,2H), 4.55(t,2H), 4.76(s,2H), 5.11(d,1H), 5.60(d,1H), 5.70(brd,1H), 6.75(d,1H), 7.03˜7.17(d,1H), 9.59(d, 1H).

EXAMPLE 16 Synthesis of7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-dimethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

p-methoxybenzyl7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-iodo-1-propen-1-yl]3-cephem-4-carboxylate(1.2g) and 4-dimethylamino-2-butenylamide(0.32 g) were reacted in the samemanner as described in Example 13 to obtain the target product(230 mg).

mp: 184° C.(dec.); IR(KBr,cm⁻¹): 1762, 1675, 1598, 1520; NMR(DMSO-d₆) δ(ppm): 3.10(s,6H), 3.88(d,2H), 3.92(d,2H), 3.93(d,2H), 4.25(t,2H),4.52(t,2H), 4.77(s,2H), 5.15(d,1H), 5.59(d,1H), 5.71(br d,1H),6.72(d,1H), 7.01˜7.22(br d,1H), 7.15˜7.34(d,1H), 9.62(d,1H).

EXAMPLE 17 Synthesis of p-methoxybenzyl7β-benzylideneamino-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylateiodide

To a solution of p-methoxybenzyl7β-benzylideneamino-3-[(Z)-3-chloro-1-propen-1-yl]3-cephem-4-carboxylate(1.6 g) dissolved in acetone(10 ml), sodium iodide(1.5 g) was addedthereto under ice-cooling. The resulting solution was stirred for 3hours at room temperature. The solvent was distilled off, and theresidue was extracted with ethyl acetate. The extract was washed with a10%-sodium thiosulfate and with a saturated brine, followed by dryingwith addition of anhydrous magnesium sulfate. The dried extract wasconcentrated to about 20 ml. To the concentrate was added4-ethylmethyl-amino-2-butenylamide (0.57 g) in ethyl acetate(30 ml). Themixture was stirred for 2 hours at 0° C., and ethyl ether(50 ml) wasadded thereto. The resulting precipitates were collected by filtrationto obtain the target product(1.5 g).

IR(KBr,cm⁻¹): 1762, 1678, 1600; NMR(DMSO-d₆) δ (ppm): 1.27(t,3H),3.14(s,3H), 3.41˜3.82(m,4H), 3.73(s,3H), 3.95(br,2H), 4.26(m,2H),5.23(d,1H), 5.44(d,1H), 5.74(br d,1H), 6.17(m,1H), 6.79(d,1H),7.01˜7.19(br d,1H), 7.28(d,1H), 7.3˜7.9(m,9H), 8.51(br s,1H).

EXAMPLE 18 Synthesis of7β-amino-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylatehydrochloride

The compound(1.5 g) prepared in Example 17 was added 90%-formic acid(4.2ml) and 35%-hydrochloric acid(0.85 ml). The resulting mixture wasstirred for 90 minutes at room temperature, and acetone(500 ml) wasadded thereto. The resulting precipitates were collected by filtrationto obtain the target product(0.6 g).

IR(KBr,cm⁻¹): 1786, 1688; NMR(DMSO-d₆) δ (ppm): 1.27(t,3H), 3.11(s,3H),3.24˜4.35(m,6H), 4.08(br,2H), 5.22(br,2H), 6.0˜6.5(m,1H), 7.04(d,1H),7.27(d,1H), 7.36(m,1H).

EXAMPLE 19 Synthesis of7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoacetamido]-3-[(E)-3-[(1-carbamoyl-1-propen-3-yl)-3-ethylmethylammonio]-1-propen-1-yl]3-cephem-4-carboxylate

The compound(0.6 g) prepared in Example 18 was added water:methylalcohol(1:6, 26 ml), sodium acetate(0.61 g) and(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-fluoroethoxyiminoaceticacidchloridehydrochloride (0.43 g). The resulting mixture was stirred for 3 hours at25˜30° C. After the mixture cooled to 0° C., insoluble materials werefiltered off. The filtrate was chromatographed oversilicagel[acetonitrile:distilled water 4:1(v/v)], Sephadex LH-20(orAmberlite XAD-2), and then lyophilized to obtain the target product(420mg) as white solid.

mp, IR, NMR: Identical with Example 14

Experimental Tests

1. in vitro activity test

In order to illustrate the surprisingly superior antibacterial activityof the compounds of the present invention, the minimal inhibitoryconcentrations(MIC) of the compounds synthesized in the above Exampleswere determined and compared with Cefotaxime and Cefpirome which wereused as the control compounds. These MIC values were determined by agardilution method: that is, two-fold dilutions of each of the testcompounds were made and dispersed in a Mueller-Hinton Broth medium.Standard test strain which had the 10₆ CFU per ml was inoculated on themedium, and was incubated at 37° C. for 18 to 20 hours. The results ofthe MIC tests are shown in Table 1.

                                      TABLE 1                                     __________________________________________________________________________    Minimal Inhibitory Concentration(MIC) of test compounds (μg/ml)            __________________________________________________________________________                     Example                                                                            Example                                                                            Example                                              Test Strain 1 6 12 CTX CPR                                                  __________________________________________________________________________     1.                                                                             Streptococcus pyogenes 77A                                                                   0.006                                                                              0.012                                                                              0.025                                                                              0.012                                                                              0.012                                       2. Streptococcus faecium MD 8b 50 100> 100> 100> 100                          3. Staphylococcus aureus SG511 0.4 0.8 0.8 1.6 0.8                            4. Staphylococcus aureus 285 0.4 0.4 0.8 1.6 0.4                              5. Staphylococcus aureus 503 0.8 0.8 0.8 3.1 0.8                              6. Escherichia coli 078 <0.006 0.12 0.025 0.025 0.025                         7. Escherichia coli DC 0 0.025 <0.006 <0.006 0.012 0.012                      8. Escherichia coli DC 2 0.05 0.05 0.05 0.05 0.1                              9. Escherichia coli TEM 0.05 0.05 0.05 0.05 0.05                             10. Escherichia coli 1507E 0.025 0.05 0.05 0.025 0.025                        11. Pseudomonas aeruginosa 9027 12.5 6.3 12.5 25 6.3                          12. Pseudomonas aeruginosa 1592E 3.1 3.1 6.3 25 3.1                           13. Pseudomonas aeruginosa 1771 6.3 3.1 6.3 12.5 3.1                          14. Pseudomonas aeruginosa 1771M 0.4 0.4 0.8 0.4 0.8                          15. Salmonella typhimurium 0.1 0.1 0.4 0.1 0.1                                16. Klebsiella aerogenes 1082E 6.3 3.1 3.1 6.3 6.3                            17. Klebsiella aerogenes 1522E 0.05 0.1 0.1 0.1 0.1                           18. Enterobacter cloace P99 0.4 1.6 1.6 100> 3.1                              19. Enterobacter cloace 1321E 0.012 0.025 0.05 0.05 0.025                   __________________________________________________________________________                     Example                                                                            Example                                                                            Example                                              Test Strain 13 14 15 CTX CPR                                                __________________________________________________________________________     1.                                                                             Streptococcus pyogenes 71A                                                                   0.025                                                                              0.05 0.025                                                                              0.012                                                                              0.012                                       2. Streptococcus faecium MD 8b 100> 100> 100> 100> 100                        3. Staphylococcus aureus SG511 0.8 0.4 0.8 1.6 0.8                            4. Staphylococcus aureus 285 0.8 0.8 0.8 1.6 0.4                              5. Staphylococcus aureus 503 0.8 0.8 0.8 3.1 0.8                              6. Escherichia coli 078 0.05 0.012 0.025 0.025 0.025                          7. Escherichia coli DC 0 0.025 <0.006 0.012 0.012 0.012                       8. Escherichia coli DC 2 0.2 0.05 0.1 0.05 0.1                                9. Escherichia coli TEM 0.05 0.025 0.05 0.05 0.05                            10. Escherichia coli 1507E 0.1 0.025 0.1 0.025 0.025                          11. Pseudomonas aeruginosa 9071 3.1 12.5 3.1 25 6.3                           12. Pseudomonas aeruginosa 1592E 1.6 3.1 3.1 25 3.1                           13. Pseudomonas aeruginosa 1771 1.6 3.1 3.1 12.5 3.1                          14. Pseudomonas aeruginosa 1771M 0.4 0.4 0.8 0.4 0.8                          15. Salmonella typhimurium 0.2 0.1 0.2 0.1 0.1                                16. Klebsiella aerogenes 1082E 3.1 3.1 3.1 6.3 6.3                            17. Klebsiella aerogenes 1522E 0.2 0.025 0.05 0.1 0.1                         18. Enterobacter cloace P99 1.6 0.4 0.8 100> 3.1                              19. Enterobacter cloace 1321E 0.05 0.012 0.05 0.05 0.025                    __________________________________________________________________________      CTX: Cefotaxime                                               CPR: Cefpirome                                          

As can be seen from Table 1, the cephalosporin compounds of the presentinvention possess potent and broad antibacterial activities as comparedwith the known broad spectrum cephalosporin antibiotics, Cefotaxime andCefpirome. More specifically, MICs of Example 1, 6, 12, and 14 againstStaphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa weresuperior to that of Cefotaxime and similar to that of Cefpirome. Also,MICs of Example 13 against Pseudomonas aeruginosa was superior to thatof Cefpirome.

2. Acute toxicity studies

In order to illustrate usefulness of the compounds of the presentinvention, acute toxicity test of the compounds synthesized in the aboveExamples were carried out. Each dose of the compounds dissolved insaline and buffer solution (PH=7.0) was administered by intravenous orsubcutaneous injection. Mortalities of the animals was recorded sevendays latter. The results of the acute toxicity studies are shown inTable 2.

                  TABLE 2                                                         ______________________________________                                                         LD.sub.50 mg/kg                                              Compound         i.v     s.c                                                  ______________________________________                                        Example 1        >3000   >4000                                                  Example 2 >2000 >3000                                                         Example 6 >3000 >3000                                                         Example 12 >2000 >3000                                                        Example 13 >2000 >3000                                                        Example 15 >2000 >3000                                                      ______________________________________                                          Mouse: Male ICR strain, 4 weeks                         

The compounds of the Example 1, 2, 6, 12, 13 and 15 are shown highstability as an antimicrobial medicament from >2000 mg/Kg in intravenousroutes and >3000 mg/Kg in subcutaneous routes. Accordingly, thecompounds of the present invention can be used in the therapeutictreatment of human beings or animals infected with variety ofGram-positive or Gram-negative bacteria.

The compounds of the present invention may be administered 1˜3 times inan amount ranging from 50˜5000 mg/day. The present invention alsoincludes within its scope pharmaceutical compositions comprising one ormore of the compound(I) and their derivatives as active ingredients, inassociation with pharmaceutically acceptable carriers, excipients orother additives, if necessary. The compositions may be formulated intovarious forms such as tablets, capsules, solution, injection, syrup,ointment, cream, suppositories, which may contain conventional additivessuch as a dispersant, suspending agent, stabilizer and the like.

Formulation examples are described below.

    ______________________________________                                        Formulation example 1                                                         ______________________________________                                        The compound of prepared Example 1                                                                  100 mg                                                    Lactose 100 mg                                                                Corn starch  50 mg                                                            Talc  45 mg                                                                   Magnesium stearate  5 mg                                                    ______________________________________                                    

The above compositions are formulated into tablets by the conventionaltablets preparation method.

    ______________________________________                                        Formulation example 2                                                         ______________________________________                                        The compound of prepared Example 6                                                                  100 mg                                                    Lactose  50 mg                                                                Corn starch 100 mg                                                            Talc  20 mg                                                                   Magnesium stearate  5 mg                                                    ______________________________________                                    

The above compositions are formulated into capsules by the conventionalcapsules preparation method.

    ______________________________________                                        Formulation example 3                                                         ______________________________________                                        The compound of prepared Example 13                                                                  250 mg                                                   Sodium chloride 12.5 mg                                                     ______________________________________                                    

The above compositions are formulated into injection by the conventionalinjection preparation method.

What is claimed is:
 1. A compound of the following formula (I) or apharmaceutically acceptable non-toxic salt thereof: ##STR6## whereinR_(1a) is a hydrogen atom; R_(1b) is a hydrogen atom, C₁₋₄ alkyl groupor fluoro-substituted C₁₋₃ alkyl group;Q is CH or N; R₂ and R₃ may bethe same or different and are individually a C₁₋₃ alkyl group; and R₄ is##STR7## wherein R₇ is a C₁₋₃ alkyl group; Y is O or S; andZ is ahydrogen atom or C₁₋₃ alkyl group.
 2. The compound of formula (I) or thepharmaceutically acceptable salt thereof recited in claim 1, whereinR_(1a) is a hydrogen atom; R_(1b) is a methyl group, ethyl group,fluoromethyl group or 2-fluoroethyl group;R₂ and R₃ may be the same ordifferent and are individually a methyl or ethyl group; Q is CH or N;and R₄ is ##STR8## wherein R₇ is a methyl group; Y is O or S; and Z is ahydrogen.
 3. A compound of the following formula (VIII): ##STR9##wherein R₂ and R₃ may be the same or different and are individually aC₁₋₃ alkyl group;X is a halogen atom; and R₄ is ##STR10## wherein R₇ isa C₁₋₃ alkyl group; Y is O or S; andZ is a hydrogen atom or C₁₋₃ alkylgroup.
 4. A pharmaceutical composition comprising a therapeuticallyeffective amount of the cephalosporin compound (I) or thepharmaceutically acceptable non-toxic salt thereof: ##STR11## whereinR_(1a) is a hydrogen atom; R_(1b) is a hydrogen atom, C₁₋₄ alkyl groupor fluoro-substituted C₁₋₃ alkyl group;Q is CH or N; R₂ and R₃ may bethe same or different and are individually a C₁₋₃ alkyl group; and R₄ is##STR12## wherein R₇ is a C₁₋₃ alkyl group;Y is O or S; and Z is ahydrogen atom or C₁₋₃ alkyl group.